Uro-genital condition treatment system

ABSTRACT

The present invention is directed to a system for treating uro-genital conditions. One aspect of this invention involves the treatment system comprising one or more polypeptides with a amino acid sequence including KPV (SEQ. ID. NO. 1), MEHFRWG (SEQ. ID. NO. 2), HFRWGKPV (SEQ. ID. NO. 3), SYSMEHFRWGKPV (SEQ. ID. NO. 4), for treatment of uro-genital conditions. The one or more polypeptides can also be a dimer formed from any of the amino acid sequence above. Uro-genital conditions can include infections, inflammation, or both. In one preferred embodiment of the invention, the uro-genital condition includes infection and/or inflammation of the vagina, vulva, urinary tract, penis, and/or the rectum. In another preferred embodiment of the invention, the one or more polypeptides are dissolved in a carrier. In another preferred embodiment of the invention, the one or more polypeptides are associated with a tampon for preventing toxic shock syndrome. In another preferred embodiment, the one or more polypeptides are associated with a contraceptive for prevention of sexually transmitted diseases or infections. In another preferred embodiment, the one or more polypeptides are associated with a suppository for insertion into the vagina or rectum.

[0001] The present application is a Divisional of U.S. application Ser.No. 09/535,066, filed Mar. 23, 2000 and now pending, which claimspriority to U.S. Provisional Patent Application Serial No. 60/126,233filed Mar. 24, 1999.

FIELD OF THE INVENTION

[0002] The present invention relates to the field of treatment foruro-genital conditions.

BACKGROUND OF THE INVENTION

[0003] Uro-genital conditions or diseases commonly affect both men andwomen. These conditions include infections and/or inflammation of theurinary system and the genital system. For example, according to theNational Institute of Child Health and Human Development (NICHD), “mostwomen will have at least one form of vaginitis in their lifetime.”Vaginitis, National Institute of Child Health and HumanDevelopment—Publications On-line, (last modified Jan. 12, 2000),<http://www.nichd.nih.giv/publications/pubs/vag1.htm>. The causes forvaginitis range from bacterial, fungal, or viral infections toirritations from chemicals in creams, sprays, or even clothing that arein contact with this area. Id. For women with bacterial and fungalinfections, these infectious agents often originate from the rectal areaand migrate across the perineum to reach the vagina or the urethra.

[0004] A common type of vaginitis is candidiasis or yeast infection thatis most commonly caused by Candida albicans. The Candida species arepart of an individual's normal flora of microbial organisms present inskin, mouth, and the gastrointestinal tract. Robbins Pathologic Basis ofDisease 5^(th) ed., Saunders Co., Philadelphia (1994) p. 354. They alsolive in small number in a woman's vagina. They grow best in warm, moistsurfaces such as the vagina or the oral cavity. They are normallynon-pathogenic, but when a change in their environment occurs, such asin response to a woman's hormonal changes in menopause, pregnancy, or inresponse to stress, they can overgrow to cause a yeast infection. Thesechanges can also occur in immunosuppressed or compromised individualssuch as people undergoing chemotherapy, taking immunosuppresants, orafflicted with AIDS.

[0005] Current treatment for candidiasis includes over the counter drugswith active ingredients such as butoconazole nitrate (Femstat®),clotrimazole (Gyne-Lotrimin® and others), miconazole (Monistat® andothers), and tioconazole (Vagistat®). These drugs are topically appliedin the vagina and break down Candida's cell wall. Other similartreatments include prescription drugs with active ingredients in thesame family such as fluconazole (Diflucan®), terconazole (Terazol®), andketoconazole (Nizoral®).

[0006] Although vaginitis has been commonly associated with Candida,bacterial vaginosis is actually the most common vaginal infection inwomen of reproductive age according to the NICHD. Vaginitis, supra.Overgrowing of bacteria in the vagina causes bacterial vaginosis muchlike Candida, but the drugs used for its treatment are different.

[0007] On the other hand, men can also contract Candida infections ontheir penis involving the glans and the prepuce. Balanoposthitis, anonspecific infection of the glans and prepuce, is caused by a widevariety of organisms including fungi such as Candida and pyogenicbacteria such as staphylococci. Robbins Pathologic Basis of Disease5^(th) ed., Saunders Co., Philadelphia (1994) p. 1008.

[0008] Staphylococci are gram positive bacteria that are normallypresent in skin and other mucosal membranes of the body. Staphylococcusaureus, in particular, is a virulent pathogen that causes a myriad ofconditions and diseases stemming from skin lesions, endocarditis,respiratory infection, food poisoning, to toxic shock syndrome. Forwomen using highly absorbent tampons, it is known that S. aureus cancolonize the vagina and secrete a toxin called toxic shock syndrometoxin (TSST-1). According to the Food and Drug Administration,approximately half of the toxic shock syndrome cases reported today areassociated with tampon use during menstruation and usually in youngwomen. Tampons and Asbestos, Dioxin, & Toxic Shock Syndrome, FDA Centerfor Devices and Radiological Health (Jul. 23, 1999),<http://www.fda.gov/cdrh/ocd/tamponsabs.html>.

[0009]S. aureus infections are commonly treated with methicillin.Although it is very effective, some strains of S. aureus have developedresistance to methicillin, and only a few antibiotics can successfullytreat these methicillin-resistant Staphylococcus aureus (MRSA). One ofthese antibiotics commonly used for MRSA is vancomycin. A strain of S.aureus, however, with reduced susceptibility to vancomycin (VISA) hasalready been identified. Khurshid, M. A., et. al., Staphylococcus aureuswith Reduced Susceptibility to Vancomycin—Illinois 1999, Morbidity andMortality Weekly Report, 48(51): 1165-1167 (2000),<http://www.cdc.gov/epo/mmwr/preview/mmwrhtml/mm4851a1.htm>. Theemergence of antibiotic resistant bacterial strains has created a needfor alternative ways to combat bacterial infections.

[0010] In addition to infection by fungi and bacteria, viral vaginitisis also common. These infections are most often transmitted throughsexual intercourse. Viral vaginitis includes infection by herpes simplexvirus (HSV) or human papillomavirus (HPV). HSV viruses, for example,replicate in the genital area, which is the site of entrance, and alsoinfect the neurons that innervate the genitals. To avoid the body'simmune system, HSV viruses can remain latent in these neurons, andbecome reactivated in response to environmental conditions such asstress, immunosuppression, irradiation, or viral infection. Currenttreatments for HSV include drugs such as acyclovir, famciclovir, orvalacyclovir.

[0011] As for the urinary system, according to the American MedicalAssociation, urinary tract infections (UTIs) are one of the most commondisorders prompting a physician visit. Women's Health, Urinary TractInfections: A Patient's Guide to Treatment, AMA Health Insight, On-LineHealth Information for Everyone (last updated Oct. 30, 1998)<http://www.amaassn.org/insight/h_focus/wom_hlth/uti/uti.htm>. Theseinfections are most often caused by Escherichia coli, but can alsoinvolve organisms such as Candida and Staphylococci. Id. Theseinfections can start at the urethra and travel up to the bladder causingcystitis. Ultimately, it can even ascend to the kidneys through theureters and cause pyelonephritis. Both men's and women's urinary systemscan become infected with these microorganisms.

[0012] Since uro-genital conditions are not confined to one singlecause, current treatments require different drugs to treat specificcauses. These causes have to be first identified. Identificationrequires time, but more so, requires a gynecological examination forwomen to determine the specific infectious agents or lack thereof.

[0013] With the increased use of antibiotics and other drugs,microorganisms, such as methicillin-resistant staphylococcus aureus, areincreasingly developing resistance to currently available drugs. Thus, acontinuing need exists for new classes of drugs that can combat thebroad spectrum of infectious agents.

SUMMARY OF THE INVENTION

[0014] The present invention is directed to a system for treatinguro-genital conditions. One aspect of this invention involves thetreatment system comprising one or more polypeptides with a amino acidsequence including KPV (SEQ. ID. NO. 1), MEHFRWG (SEQ. ID. NO. 2),HFRWGKPV (SEQ. ID. NO.3), or SYSMEHFRWGKPV. (SEQ. ID. NO. 4) fortreatment of uro-genital conditions. The one or more polypeptides canalso be a dimer formed from any of the-amino acid sequence above.Uro-genital conditions can include infections, inflammation, or both. Inone preferred embodiment of the invention, the uro-genital conditionincludes infection and/or inflammation of the vagina, vulva, urinarytract, penis, and/or the rectum. In another preferred embodiment of theinvention, the one or more polypeptides are dissolved in a carrier. Inanother preferred embodiment of the invention, the one or morepolypeptides are associated with a tampon for preventing toxic shocksyndrome. In another preferred embodiment, the one or more polypeptidesare associated with a contraceptive for prevention of sexuallytransmitted diseases or infections. In another preferred embodiment, theone or more polypeptides are associated with a suppository for insertioninto the vagina or rectum. In another preferred embodiment of theinvention, the one or more polypeptides are dissolved in a liquidcarrier for douching the vagina.

BRIEF DESCRIPTION OF THE DRAWINGS

[0015]FIG. 1 shows the inhibitory effects of α-MSH and/or itsderivatives on the growth of S. aureus.

[0016]FIG. 2 shows the effects of α-MSH and/or its derivatives onurokinase induced growth of S. aureus.

[0017]FIG. 3 shows the inhibitory effects of α-MSH and/or itsderivatives on the growth of C. albicans.

[0018]FIG. 4 compares the anti-fungal activities of α-MSH and/or itsderivatives with fluconazole.

[0019]FIGS. 5A to 5D show the inhibitory effects of α-MSH and/or itsderivatives on C. albicans' germ tube formation.

[0020]FIG. 6 shows the enhanced neutrophil-killing effects of α-MSHand/or its derivatives against C. albicans.

[0021]FIGS. 7, 8, and 9 show the mechanism by which α-MSH and/or itsderivatives inhibit the growth of C. albicans.

[0022] FIGS. 10-13 show the inhibitory effects of α-MSH and/or itsderivatives on viral replication, expression in chronically infectedcells.

[0023]FIG. 14 shows the mechanisms by which α-MSH and/or its derivativesinhibit viral replication, expression, and reactivation.

[0024]FIG. 15 shows the inhibitory effects of α-MSH and/or itsderivatives on viral replication, expression in acutely infected cells.

[0025]FIG. 16 shows a representation of the chemical structure of oneform of the KPV dimer for use with one aspect of the invention.

GENERAL DESCRIPTION OF THE INVENTION

[0026] The references cited below are hereby incorporated by referenceas if fully set forth herein. The present invention involves a methodand system for treating uro-genital conditions with the use ofalpha-melanocyte stimulating hormone (“α-MSH”) and/or its derivatives.α-MSH is an ancient thirteen amino-acid peptide (SEQ. ID. NO. 4)produced by post-translational processing of the larger precursormolecule propiomelanocortin. It shares the 1-13 amino acid sequence withadrenocorticotropic hormone (“ACTH”), also derived frompropiomelanocortin. α-MSH is known to be secreted by many cell typesincluding pituitary cells, monocytes, melanocytes, and keratinocytes. Itcan be found in the skin of rats, in the human epidermis, or in themucosal barrier of the gastrointestinal tract in intact andhypophysectomized rats. See e.g. Eberie, A. N., The Melanotrophins,Karger, Basel, Switzerland (1998); Lipton, J. M., et. al.,Anti-inflammatory Influence of the Neuroimmunomodulator α-MSH, Immunol.Today 18, 140-145 (1997); Thody, A. J., et.al., MSH Peptides are Presentin Mammalian Skin, Peptides 4, 813-815 (1983); Fox, J. A., et.al.,Immunoreactive α-Melanocyte Stimulating Hormone, Its Distribution in theGastrointestinal Tract of Intact and Hypophysectomized Rats, Life. Sci.18, 2127-2132 (1981).

[0027] α-MSH and its derivatives are known to have potent antipyreticand anti-inflammatory properties, yet they have extremely low toxicity.They can reduce production of host cells' proinflammatory mediators invitro, and can also reduce production of local and systemic reactions inanimal models for inflammation. The “core” α-MSH sequence (4-10) (SEQ.ID. NO. 2), for example, has learning and memory behavioral effects butlittle antipyretic and anti-inflammatory activity. In contrast, theactive message sequence for the antipyretic and anti-inflammatoryactivities resides in the C-terminal amino-acid sequence of α-MSH, thatis, lysine-proline-valine (“Lys-Pro-Val” or “KPV”) (SEQ. ID. NO. 1).This tripeptide has activities in vitro and in vivo that parallel thoseof the parent molecule. The anti-inflammatory activity of α-MSH and/orits derivatives are disclosed in the following two patents and arehereby incorporated by reference: U.S. Pat. No. 5,028,592, issued onJul. 2, 1991 to Lipton, J. M., entitled Antipyretic andAnti-inflammatory Lys Pro Val Compositions and Method of Use; U.S. Pat.No. 5,157,023, issued on Oct. 20, 1992 to Lipton, J. M., entitledAntipreytic and Anti-inflammatory Lys Pro Val Compositions and Method ofUse; see also Catania, A., et. al., α-Melanocyte Stimulating Hormone inthe Modulation of Host Reactions, Endocr. Rev. 14, 564-576 (1993);Lipton, J. M., et.al., Anti-inflammatory Influence of theNeuroimmunomodulator of α-MSH, Immunol. Today 18, 140-145 (1997);Rajora, N., et. al., α-MSH Production Receptors and Influence onNeopterin, in a Human Monocyte/macrophage Cell Line, J. Leukoc. Biol.59, 248-253 (1996); Star, R. A., et. al., Evidence of AutocrineModulation of Macrophage Nitric Oxide Synthase by α-MSH, Proc. Nat'l.Acad. Sci. (USA) 92, 8015-8020 (1995); Lipton, J. M., et.al.,Anti-inflammatory Effects of the Neuropeptide α-MSH in Acute Chronic andSystemic inflammation, Ann. N.Y. Acad Sci. 741, 137-148 (1994); Fajora,N., et.al., α-MSH Modulates Local and Circulating tumor Necrosis Factorα in Experimental Brain Inflammation, J. Neuroosci, 17, 2181-2186(1995); Richards, D. B., et. al., Effect of α-MSH (11-13)(lysine-proline-valine) on Fever in the Rabbit, Peptides 5, 815-817(1984); Hiltz, M. E., et. al., Anti-inflammatory Activity of aCOOH-terminal Fragment of the Neuropeptide α-MSH, FASEB J. 3, 2282-2284(1989).

[0028] In addition to its anti-inflammatory and anti-pyretic function,one aspect of the present invention involves the anti-microbial oranti-infection activity of α-MSH and/or its derivatives. As describedbelow, α-MSH and/or its derivatives have significant anti-infectionuses, including, for example, use in reducing the viability of microbes,reducing the germination of yeast, killing microbes without reducing thekilling of microbes by human neutrophils, for treating inflammationassociated with microbial infection without reducing microbial killing,increasing the accumulation of cAMP in microbes, and inhibiting thereplication and expression of viral pathogens.

[0029] In a preferred embodiment of the invention, these anti-microbialor anti-infection activities are most particularly associated with theC-terminal amino-acid sequence—KPV. This tripeptide, along with α-MSHand its derivatives, are effective over a very broad range ofconcentrations, including picomolar concentrations that normally occurin human plasma.

[0030] As discussed in the background section, uro-genital conditionsare not confined to one single cause. Multiple organisms and infectiousagents, from, bacteria, fungi, to viruses, individually or incombination can cause a wide variety of conditions including vaginitis,vulvitis, urethritis, balanophosthithis, candidiasis, sexuallytransmitted diseases, and toxic shock syndrome. For treatment of theseconditions, α-MSH and/or its derivatives can be applied locally to thesite of the infection and/or inflammation by methods known in the art.For example, α-MSH and its derivatives can be dissolved in solutionssuch as phosphate buffer saline, hyalurinate, methylcellulose,carboxymethlcellulose, or ethanol. Common carriers such as ointment,cream, gel, dissolvable pill, aerosol spray, suppository, liquidsolution for douche, or the absorbent material of tampons can carryα-MSH and/or its derivatives as active ingredients for treatinguro-genital conditions. These carriers can be applied to the site of theinfection or inflammation by an applicator such as syringes orsyringe-like apparati, bandages, catheters, tubes with a plunger,spatula or other types of flat surface applicators, condoms, sponges,diaphragms, tampon applicators, or fingers.

[0031] More specifically, the preferred embodiment of the invention isto dissolve α-MSH and/or its derivatives in a liquid-based carrier. Thiscarrier carrying the solvated α-MSH and/or its derivatives is thenstored in a pressurized canister. Upon release of the carrier by arelease valve or other mechanisms from the pressurized canister, anaerosol foam is formed and captured into a syringe or a syringe-typeapparatus. The syringe is then partially inserted into the vagina andits contents delivered into the vaginal canal. The syringe orsyringe-type apparatus and its opening can also be molded to differentsize, shapes, and lengths to accommodate insertion into differenturo-genital areas such as the urethra or the rectum.

[0032] Another preferred embodiment of the invention is a suppositorythat comprises a carrier. This carrier such as a gel or glycerin issolid or semi-solid at room temperature, but melts at body temperaturewhen inserted into the vagina or rectum. This carrier carrying thesolvated α-MSH and/or its derivatives are delivered into the site ofuro-genital condition when the carrier melts.

[0033] Delivering α-MSH and its derivatives to the outside area of theuro-genital area such as the vulva or the glans and prepuce of the peniscan be achieved by topically applying a cream, ointment, gel, spray,foam, or balm, the compositions of which are already well known in theart.

[0034] In another aspect of the invention, tampons can be treated withα-MSH and/or its derivatives during the manufacturing process. Thepresence of α-MSH in tampons may inhibit the growth of microorganismssuch as Staphylococcus aureus that secretes the toxic shock syndrometoxin (TSST-1). The processes for making tampons are already well knownin the art. Treatment of the tampon's absorbent material with α-MSH orits derivatives may be accomplished by first soaking the absorbentmaterial in a solution of α-MSH and/or its derivatives. The absorbentmaterial can then be allowed to dry. Alternatively, α-MSH may besprinkled onto the tampon's absorbent material as dry powder.

[0035] In another aspect of the invention, α-MSH and/or its derivativesmay be delivered to the site of the infection by using contraceptivessuch as condoms, diaphragms, sponges, or other barrier-type mechanismsused for preventing pregnancy or sexually transmitted diseases. α-MSHand/or its derivatives can be dissolved in the lubricant used incondoms, in the gel or foam used together with the diaphragms, or in anyother spermicidal solution used in conjunction with condoms, diaphragms,or sponges.

[0036] In another aspect of the invention, α-MSH and/or its derivativesmay be dissolved in a liquid for use with a douche. The liquid can bedelivered by the douche into the vagina for treating infection and/orinflammation.

[0037] The following examples demonstrate the ability and application ofα-MSH and its derivatives to combat infection. Methods in microbiology,molecular biology, and cell culture used but not explicitly described inthis disclosure have already been amply reported in the scientificliterature. These methods are well within the ability of one skilled inthe art.

[0038] The peptides used in the following examples include: α-MSH (1-13)(SEQ. ID. NO. 4), (4-10) (SEQ. ID. 002), (6-13) (SEQ. ID. 003), and(11-13) (SEQ. ID. NO. 1), all of which were N-acetylated and C-amidated,and ACTH (1-39) and (18-39) (CLIP). These peptides were prepared bysolid-phase peptide synthesis and purified by reversed phased highperformance liquid chromatography. Some examples also include a dimer ofthe amino acid sequence CKPV (SEQ. ID. NO. 8), which also wasN-acetylated and C-amidated (the “KPV dimer”). FIG. 16 shows arepresentation of the chemical structure for this KPV dimer. Dimers canbe formed by adding cysteines at the N-termini of any of the abovepolypeptides and allowing the cysteines of two polypeptides to form adisulfide bond. Both homo-dimers and hetero-dimers can be formed usingthis method.

[0039] Statistical significance disclosed in the examples below wasanalyzed using one-way analysis of variance and the Student's t test.Probability values greater than 0.05 were considered significant.

EXAMPLE I

[0040] This example illustrates the anti-microbial properties of α-MSHand/or its derivatives against Staphylococcus aureus.

[0041] Cultures of S. aureus (ATCC 29213) were obtained from thecollection of the Department of Microbiology, Ospedale Maggiore diMilano. S. aureus (1×10⁶/ml in Hank's balanced salt solution) wasincubated in the presence or absence of α-MSH (1-13) (SEQ. ID. NO. 4),α-MSH (11-13) (SEQ. ID. NO. 1), or the KPV dimer at concentrations inthe range of 10⁻¹⁵ to 10⁻⁴ M for two hours at 37° C. Cells were thenwashed in cold distilled water and diluted with HBSS to a concentrationof 100 organisms/ml. One-milliliter aliquots were dispensed on bloodagar plates and incubated for 24 hours at 37° C. Viability of themicroorganisms was estimated from the colonies formed. In another set ofexperiments, 500 units of urokinase, a S. aureus growth enhancer, werealso incubated with the bacteria (10⁵/100 ml) for four hours at 37° C.in a shaking water bath together with the peptides.

[0042]FIG. 1 shows that α-MSH (1-13) (SEQ. ID. NO. 4), α-MSH (11-13)(SEQ. ID. NO. 1), and the KPV dimer all inhibited S. aureus colonyformation. These inhibitory effects occurred over a wide range ofconcentrations and were significant (p>0.01) with peptide concentrationsof 10⁻¹² to 10⁻⁴ M. FIG. 2 shows that α-MSH (1-13) (SEQ. ID. NO. 4) andα-MSH (11-13) (SEQ. ID. NO. 1) at concentrations of 10⁻⁶ M significantlycountered the growth enhancing effect of urokinase. Thus, α-MSH or itsderivatives can inhibit the growth of Staphylococcus aureus, an agentknown to cause toxic shock syndrome associated with tampon use,vaginitis, UTIs, urethritis, and balanoposthitis.

EXAMPLE II

[0043] This example illustrates the anti-fungal properties of α-MSHand/or its derivatives against Candida albicans.

[0044] Clinical isolates of C. albicans were also obtained from thecollection of the Department of Microbiology, Ospedale Maggiore diMilano. Cultures of C. albicans were maintained on Sabouraud's agarslants and periodically transferred to Sabouraud's agar plates andincubated for 48 hours at 28° C. To prepare stationary growth-phaseyeast, a colony was taken from the agar plate, transferred into 30 ml ofSabouraud-dextrose broth, and incubated for 72 hours at 32° C. Cellswere centrifuged at 1000×g for ten minutes, and the pellet was washedtwice with distilled water. Cells were counted and suspended in Hank'sbalanced salt solution (“HBSS”) to the desired concentration. Viability,determined by exclusion of 0.01% methylene blue, remained greater than98%.

[0045] At 1×10⁶/ml in HBSS, these fungi were incubated in the presenceor absence of α-MSH (1-13) (SEQ. ID. NO. 4), α-MSH (11-13) (SEQ. ID. NO.1), or the KPV dimer at concentrations ranging from 10⁻¹⁵ to 10⁻⁴ M fortwo hours at 37° C. Cells were then washed in cold distilled water anddiluted with HBSS to a concentration of 100 organisms/ml. One-milliliteraliquots were then dispensed on blood agar plates and incubated for 48hours at 37° C. The organism's viability was estimated from the numberof colonies formed.

[0046]FIG. 3 shows that α-MSH(1-13) (SEQ. ID. NO. 4), α-MSH(11-13) (SEQ.ID. NO. 1), and the KPV dimer greatly reduced the ability of C albicansto form colony at concentrations ranging from 10⁻¹² to 10⁻⁴ M (p<0.01vs. control). Thus, this demonstrates that α-MSH or its derivatives caninhibit the growth of Candida albicans, an agent known to causecandidiasis, vaginitis, urethritis, and balanoposthitis.

EXAMPLE III

[0047] This example compares the anti-infection activities of α-MSHand/or its derivatives to fluconazole, an established anti-fungal agent.

[0048] α-MSH (1-13) (SEQ. ID. NO. 4), (4-10) (SEQ. ID. NO. 2), (6-13)(SEQ. ID. NO. 3), (11-13) (SEQ. ID. NO. 1), ACTH (1-39), (18-39), andfluconazole, at concentrations of 10⁻⁶ to 10⁻⁴ M, were tested against C.albicans using the same procedures as in Example II. FIG. 4 shows thatcompared with fluconazole, α-MSH (11-13) (SEQ. ID. NO. 1), (6-13) (SEQ.ID. NO. 3), and (1-13) (SEQ. ID. NO. 4) were most effective against C.albicans. Their inhibitory activities were similar to fluconazole at thesame molar concentration. In contrast, the “core” α-MSH sequence (4-10)(SEQ. ID. NO. 2), which has behavioral effects but littleanti-inflammatory activity, caused approximately 50% inhibition ofcolony forming units (CFU). Although this inhibitory effect wassubstantial (p<0.01 vs. control), it was significantly less potent thatα-MSH fragments bearing the KPV signal sequence, i.e. α-MSH (6-13) (SEQ.ID. NO. 3) and (11-13) (SEQ. ID. NO. 1) (p<0.01), or the parent moleculeα-MSH (1-13) (p<0.05). FIG. 4 also shows that ACTH (1-39) and the ACTHfragment (18-39) did not reduce C. albicans viability. Even at a higherconcentration of 10⁻⁴ M, which is not shown in the figures, ACTHpeptides were likewise ineffective.

[0049] Thus, this example demonstrates that α-MSH or its derivatives areas effective as fluconazole in inhibiting Candida's growth.

EXAMPLE IV

[0050] This example illustrates that α-MSH and its derivatives inhibitthe germination or germ tube formation of C. albicans. Germ tubeformation is a significant part of the pathogenesis of C. albicansinfection. This pathogenesis involves adhesion to host epithelial andendothelial cells and morphologic switching from the ellipsoidblastospore to various filamentous forms, e.g. germ tubes, pseudohyphae,and hyphae. Gow, N. A., Germ Tube Growth of Candida albicans, Curr.Topics Med. Myco. 8, 43-55 (1997).

[0051]C. albicans from stationary phase cultures were washed twice withdistilled water and suspended in HBSS to a final concentration of2×10⁶/ml. Hyphal growth was induced by addition of 10% inactivated horseserum (GIBCO/BRL, Paisley, Great Britain) to yeast incubated for 45minutes at 37° C. with continuous shaking. Horse serum was then removedby washing cells twice with HBSS, and incubation was further continuedfor 60 minutes at 37° C. in the presence of α-MSH (1-13) (SEQ. ID. NO.4), (6-13) (SEQ. ID. NO. 3), or (11-13) (SEQ. ID. NO. 1) at aconcentration of 10⁻⁶ M with continuous shaking. The percentage offilamentous cells was evaluated under a light microscope with the aid ofa hemocytometer. Experiments were run in triplicates and at least 200cells were scored. Photomicrographs were taken with a MC100 cameraattached to an Axioskop Zeiss microscope.

[0052]FIGS. 5A to 5D show that co-incubation of C. albicans with α-MSH(1-13) (SEQ. ID. NO. 4) or (11-13) (SEQ. ID. NO. 1)inhibited germ tubeformation induced by horse serum. α-MSH (1-13) (SEQ. ID. NO. 4) caused28-32% reduction in the number of filamentous cells while α-MSH (11-13)(SEQ. ID. NO. 1)caused 54-58% reduction. Although not shown in thefigures, α-MSH (6-13) (SEQ. ID. NO. 3)similarly had approximately 50%reduction in the number of filamentous cells. Thus, this demonstratesα-MSH or its derivatives can inhibit one mode of Candida pathogenesis byinhibiting its germ tube formation.

EXAMPLE V

[0053] Reduced killing of pathogens is a dire consequence of therapywith corticosteriods and other nonsteriodal anti-inflammatory drugsduring infection. Stevens, D. L., Could Nonsteriodal Anti-inflammatoryDrugs (NSAIDs) Enhance Progression of Bacterial Infections to ToxicShock Syndrome?, Clin. Infect. Dis. 21, 977-80 (1997); Capsoni, F., et.al., Effect of Corticosteriods on Neutrophil Function: Inhibition ofAntibody-dependent Cell-Mediated Cytotoxicity (ADCC), J.Immunopharmacol. 5, 217-30 (1983). This example illustrates that α-MSHand/or its derivatives inhibit the growth of infectious agents withoutcomprising the ability of human neutrophils to combat these infections.This example further shows that α-MSH or its derivatives can actuallyenhance the ability of neutrophils to kill these infectious agents.

[0054] Venous blood (20 ml) from healthy volunteers was anticoagulatedwith heparin. Neutrophils were then isolated using dextran sedimentationand centrifugation with Ficoll-Hypaque (Sigma Chemical Co., St. Louis,Mo., USA). Erythrocytes were lysed via hypotonic shock with theresulting neutrophils representing at least 97% of the cell suspension.Cell viability, estimated by trypan blue exclusion, was greater than98%. Neutrophils were resuspended in HBSS for the experiments.

[0055]C. albicans (1×10⁶) were opsonized with human AB serum in ashaking water bath for 30 minutes at 37° C. They were then incubatedwith neutrophils in the presence of medium alone, or medium with α-MSH(1-13) (SEQ. ID. NO. 4)or α-MSH (11-13) (SEQ. ID. NO. 1) inconcentrations ranging from 10⁻¹⁵ to 10⁻⁴ M in a shaking water bath fortwo hours at 37° C. After incubation, the culture tubes were placed onice to stop growth, and extracellular organisms were washed twice withcentrifugation at 1000-×g at 4° C. A 2.5% sodium deoxycholate solutionwas added to the suspension, and the tubes were shaken for five minutes.Cold distilled water was then added to obtain a suspension of 10⁶cells/ml. Two 1/100 serial dilution in HBSS were made to obtain a finalsuspension of 100 cells/ml. One-milliliter aliquots were then dispensedon blood agar plates and incubated for 48 hours at 37° C. Colony formingunits were counted at the end of the incubation period with experimentsrunning in triplicates and repeated using blood from five differentdonors.

[0056]FIG. 6 shows that α-MSH (1-13) (SEQ. ID. NO. 4)and (11-13) (SEQ.ID. NO. 1) actually enhanced neutrophil killing of C. albicans whenadministered at concentrations ranging from 10⁻¹² to 10⁻⁴ M (p<0.01). Itshows that this enhanced killing occurred over a very broadconcentration range including picomolar concentration, which is equal tothe concentration of α-MSH found in human plasma.

[0057] Thus, this example demonstrates that α-MSH or its derivatives cansimultaneously combat against infection and inflammation, which may alsobe applied to candidiasis, vaginitis, urethritis, balanoposthitis, orhemorrhoids.

EXAMPLE VI

[0058] This example suggests the cellular mechanism by which α-MSHand/or its derivatives exerts its anti-microbial properties in general,and anti-fungal properties in particular.

[0059]C. albicans (10⁶/ml), permeabilized with toluene/ethanol, wereincubated at 37° C. with continuous shaking in the presence or absenceof 10⁻⁶ M α-MSH (1-13) (SEQ. ID. NO. 4), (11-13) (SEQ. ID. NO. 1), orforskolin, an agent known to increase intracellular cAMP. The reactionswere stopped after three minutes by the addition of ice cold ethanol.cAMP levels were measured in duplicates using a commercial enzymeimmunoassay kit (Amersham, United Kingdom) after extraction via theliquid-phase method according to the manufacturer's instructions. In arelated experiment, C. albicans were also exposed to dideoxyadenosine(ddAdo, Sigma), a potent inhibitor of adenylyl cyclase, atconcentrations of 25, 50, and 100×10⁻⁵ M for two hours and to α-MSH orits derivatives for two additional hours. The effects of forskolin andddAdo on the ability of C. albicans to form colonies were determinedaccording to the procedures described in Example II.

[0060]FIG. 7 shows that α-MSH (1-13) (SEQ. ID. NO. 4) and (11-13) (SEQ.ID. NO. 1) enhanced cAMP content in the C. albicans. This cAMP increasewas of the same order of magnitude as that induced by equimolarforskolin. FIG. 8 shows that along with α-MSH (1-13) (SEQ. ID. NO. 4)and (11-13) (SEQ. ID. NO. 1), forskolin also significantly inhibited thegrowth of C. albicans relative to control (p<0.01). FIG. 9 shows thatddAdo has the ability to reverse the effect of α-MSH (1-13) (SEQ. ID.NO. 4) and (11-13) (SEQ. ID. NO. 1) on the growth of C. albicans.

[0061] This example demonstrates that α-MSH and its derivatives mostlikely inhibit growth of C. albicans and other microorganisms byincreasing its cAMP level, which in turn inhibits mRNA and proteinsynthesis. see e.g. Bhattacharya A., et. al., Effect of Cyclic AMP onRNA and Protein Synthesis in Candida albicans, Biochem, Biophysics. Res.Commun., 77: 1483-44 (1977).

EXAMPLE VII

[0062] This example illustrates the ability of α-MSH or its derivativesto inhibit viral replication in human cells. More specifically, α-MSHinhibited the replication and expression of HIV-1 in chronicallyinfected human monocytes.

[0063] Chronically HIV-1 infected promonocytic U1 cell line is an invitro model of latent HIV infection in monocytes. These cells carry twointegrated proviral copies of HIV, and constitutive expression of HIV isvery low. Viral replication, however, as measured by RNA transcription,p24 antigen, or reverse transcriptase release, can be activated withdifferent stimuli such as TNF-α, IL-6, IL-10, PMA or crowding of cells.

[0064] To determine the effects of α-MSH and/or its derivatives on HIVreplication, these cells were maintained in log phase of growth incomplete culture medium (RPMI 1640 supplemented with 10 mM Hepes), 2 mML-glutamine (Sigma-Aldrich), 10% heat-inactivated FCS (HycloneLaboratories, Logan, Utah, USA), penicillin at 100 units/ml andstreptomycin at 100 μg/ml (Gibco Laboratories, Grand Island, N.Y.) inlog phase of growth. Pilot experiments were first performed to determineoptimal cell density, stimuli concentration, and kinetics of HIV-1 p24antigen production using these culture conditions. Before use, cellswere washed three times with HBSS to remove extracellular viruses. Cellswere then plated onto 24-well flat-bottomed plates at a concentration of2×10⁵/ml (final volume of one ml) with medium alone or plus TNF-α (10ng/ml) (R&D Systems, Oxford, England, UK) in the presence or absence ofα-MSH (1-13) (SEQ. ID. NO. 4) or (11-13) (SEQ. ID. NO. 1) inconcentrations from 10⁻¹⁵ to 10⁻⁴ M.

[0065] In further experiments, α-MSH (11-13) (SEQ. ID. NO. 1)alone at10⁻⁵ M was added to U1 cells stimulated with TNF-α (10 ng/ml), IL-6 (20ng/ml), IL-10 (20 ng/ml) (R&D Systems, Oxford, England, UK), PMA (1ng/ml) (Sigma-Aldrich), or in crowding conditions. Crowding was achievedby seeding U1 cells at a density of 2×10⁵/ml and maintaining them inculture at 37° C. in 5% CO₂ without changing media for seven days.Cultures activated with cytokines or PMA were maintained for only 48hours. Supernatants were then removed by centrifugation and assayed forp24 antigen using commercially available ELISA kits from CellularProducts, Inc. in Buffalo, N.Y., USA. Reverse transcriptase releaseswere also measured using a commercially available kit, ELISA Retrosys RTassay from Innovagen, Lund, Sweden. For these experiments, addition ofα-MSH (11-13) (SEQ. ID. NO. 1)occurred on day one, and each conditionwas tested in triplicates.

[0066]FIG. 10 shows that α-MSH (1-13) (SEQ. ID. NO. 4)and (11-13) (SEQ.ID. NO. 1) significantly inhibited p24 antigen release from TNF-αstimulated U1 cells over a broad range of concentrations. The mosteffective concentration for both peptides was 10⁻⁵M, causing 52.7% and56.0% inhibition respectively. FIG. 11 shows that α-MSH (11-13) (SEQ.ID. NO. 1) also inhibited p²⁴ antigen and reverse transcriptase releasefrom U1 cells induced by IL-6, IL-10, PMA and in crowding condition. Inaddition, FIG. 12 shows that α-MSH(11-13) (SEQ. ID. NO. 1) alsoinhibited the transcription of both spliced and unspliced HIV-1 RNA inPMA stimulated U1 cells as measured by Northern Blot analysis.

[0067] Thus, this example demonstrates that α-MSH or its derivatives caninhibit transcription of viral genes through mediation of the TNF-α,IL-6, and IL-10 pathways.

EXAMPLE VIII

[0068] This example further illustrates the ability of α-MSH to inhibitviral replication and activation. More specifically, the addition of aneutralizing antibody to α-MSH in U1 cells substantially increasedp24-antigen release.

[0069] U1 cells were cultured similarly as described in Example VII.Endogenous α-MSH produced by U1 cells was blocked with an affinitypurified rabbit-anti-α-MSH antibody (Euro-Diagnostica, Malmo, Sweden)diluted 1:250 with medium. Control antibody was a rabbit IgG at the samedilution. Cells (2×10⁵/ml) treated with the anti-α-MSH or the controlantibody was coincubated with medium or PMA (1 ng/ml). After 48-hourincubation at 37° C., supernatants were separated and tested for p24antigen release. In crowding experiments with U1 cells cultured asdescribed above, the anti-α-MSH antibody or the control IgG was added onday one and the supernatants were harvested on day seven.

[0070]FIG. 13 shows that blocking α-MSH in resting, PMA induced, orcrowded U1 cells significantly increased the release of p24 antigen.This example strongly implies that viral replication is affected byα-MSH.

EXAMPLE IX

[0071] This example illustrates the mechanism by which α-MSH and/or itsderivatives inhibit viral replication and expression. More specifically,α-MSH or its derivatives inhibited TNF-α induced NF-κB activation andbinding.

[0072] To determine the level of NF-κB activity, nuclear extracts wereprepared from 20×10⁶ U1 cells (2×10⁵/ml in complete medium) stimulatedfor four hours with TNF-α (20 ng/ml) in the presence or absence of 10⁻⁵M α-MSH (11-13) (SEQ. ID. NO. 1). Cells were washed once with cold PBS,and twice with buffer A (10 mM Hepes pH 7.9, 1.5 mM MgCl₂, 10 mM KCl,0.5 mM PMSF and 0.5 mM DTT), centrifuged, and incubated for ten minuteson ice in buffer A plus 0.1% NP-40. Afterwards, the supernatants wereremoved, and the nuclear pellets were resuspended in 15 μl of buffer C(20 mM Hepes pH 7.9, 1.5 mM MgCl₂, 0.42 M KCl, 0.2 mM EDTA, 25%glycerol, 0.5 mM PMSF, and 0.5 mM DTT), incubated for 15 minutes on ice,mixed, and then centrifuged. The supernatants were diluted with 75 μl ofmodified buffer D (20 mM Hepes, pH 7.9, 0.05 mM KCl, 0.2 mM EDTA, 20%glycerol, 0.5 mM PMSF, and 0.5 mM DTT) and stored at −80° C. The bindingreaction was carried out for fifteen minutes at room temperature with 10μg of nuclear extract protein and 0.5 ng of ³²P-labelled NF-κB (30,000cpm/μl) or AP1 consensus in buffer A (12 mM Tris-HCl pH 7.8, 60 mM KCl,0.2 mM EDTA, 0.3 mM DTT), plus 10% glycerol, 2 μg/ml bovine serumalbumin and 1 μg/ml single stranded DNA (Pharmacia Biotech). Theoligonucleotides for NF-κB used in these studies were: +GAT CCA AGG GGACTT TCC GCT GGG GAC TTT CCA TG, and −GAT CCA TGG AAA GTC CCC AGC GGA AAGTCC CCT TG. Each oligonucleotide was annealed to its complementarystrand and end-labeled with ³²P-γ-ATP using polynucleotide kinase. Forthe determination of specific bands, nuclear extracts were firstincubated with 100 fold excess unlabeled probe for five minutes, beforeincubation with a labeled probe. The mixtures were then run on 5% (30:1)acrylamide gel in 1× TBE. The gels were dried and autoradiographed.

[0073]FIG. 14 shows that TNF-α greatly enhanced NF-κB binding activity,but the co-incubation of α-MSH (11-13) (SEQ. ID. NO. 1) at 10⁻⁵ Msignificantly reduced NF-κB activation. α-MSH (11-13) (SEQ. ID. NO. 1),however, did not alter NF-κB activation in resting cells. This suggeststhat α-MSH and/or its derivatives inhibit viral replication andexpression through regulation of the NF-κB binding.

[0074] Replication of viral agents often depends on the state ofactivation of infected cells and is often regulated by interactionsbetween viral and host factors. These host factors may include TNF-α andother cytokines such as the interleukins. Similar to HIV-1 infection andactivation, herpes simplex virus also become reactivated from latency inresponse to host cytokines. For example, TNF-α and IL-6, but not IL-1and IL-3, have been shown to reactivate HSV infection. Neutralization ofIL-6 with antibody against IL-6 significantly inhibited HSV reactivationwhile neutralization of interferon alpha and beta did not. see e.g.Baker, M., et. al., The Relationship between Interleukin-6 and HerpesSimplex Virus Type-1: Implications for Behavior and Immunopathology,Brain Behav. Immun. 13(3):201-11 (1999); Noisakran S., e. al.,Lymphocytes Delay Kinetics of HSV-1 Reactivation from in vitro Explantsof Latent Infected Trigeminal Ganglia, J. Neuroimmunol. 95(1-2):126-35(1999); Walev, I., et.al., Enhancement by TNF-alpha of Reactivation andReplication of Latent Herpes Simplex Virus from Trigeminal Ganglia ofMice, Arch Virol. 140(6):987-92 (1995); Domk-Optiz, I., et. al.,Stimulation of Macrophages by Endotoxin Results in the Reactivation of aPersistent Herpes Simplex Virus Infection, Scand J. Immunol. 32(2):69-75(1990); Fauci, A. S., Host Factors in the Pathogenesis of HIV-inducedDisease, Nature 384: 529 (1996).

[0075] TNF-α or infection by viruses, including HSV, can cause targeteddestruction of IκB, which in turn activates the nuclear translocation ofNF-κB. Nuclear translocation promotes NF-κB binding to DNA operators forthe transcription of a range of inflammatory agents including TNF-α,IL-6, and other cytokines. The expression of these cytokines, again,further reactivates other HSV infected cells to produce HSV viruses. seee.g. Patel, A., et. al., Herpes Simplex Type 1 Induction of PersistentNF-κB Nuclear Translocation Increases the Efficiency of VirusReplication, Virology 247(2):212-22 (1998).

[0076] Thus, by blocking NF-κB binding, α-MSH and/or its derivativesinhibit the expression of more inflammatory cytokines that canreactivate HSV. This example and Examples VII-VIII show that α-MSHand/or its derivatives inhibit viral replication, expression, andreactivation by inhibiting NF-κB binding in response to the body'scytokines or other viral infection.

EXAMPLE X

[0077] This example illustrates the ability of α-MSH and/or itsderivatives to inhibit viral replication in acutely infected humancells. More specifically, α-MSH inhibited the replication and expressionof HIV-1 in acutely infected human peripheral blood mononuclear cells(PBMCs).

[0078] PBMCs were isolated from normal donors by Ficoll-Hypaque densitygradient centrifugation. Monocytes were isolated by Percoll gradientseparation and allowed to differentiate into macrophages (MDM) incomplete medium of RPMI plus 20% FCS using 24-well tissue culture platesat 10⁶ cells/ml for seven days. MDM were infected with monocytotropicHIVBa-1 strain (1:10) overnight. The undiluted viral stock contained 10⁷infectious units/ml. After 24 hours, MDM were washed and resuspended incomplete medium, replaced three times a week, for three weeks. Reversetranscriptase releases were measured weekly post-infection using acommercially available kit, ELISA Retrosys RT assay from Innovagen,Lund, Sweden. 10⁻⁵ M α-MSH (11-13) (SEQ. ID. NO. 1) was added at thetime of HIV infection and daily until harvests.

[0079]FIG. 15 shows that α-MSH significantly inhibited reversetranscriptase release in acutely infected MDM. This inhibitory effectwas more pronounced on day six but was still statistically significanton day 21.

[0080] Thus, this example demonstrates that viral replication at thesite of infection can be inhibited by α-MSH or its derivatives.Consequently, the sexual transmission of venereal diseases in general,and HIV in particular can be inhibited by associating α-MSH and/or itsderivatives with contraceptives such as condoms, diaphragms, or spongesused during sexual contact, and/or the post sexual contact applicationof suppositories, cream, ointment, gel, or aerosol foams containingα-MSH and/or its derivatives

EXAMPLE XI

[0081] This example illustrates the biological functional equivalents ofα-MSH and/or its derivatives.

[0082] Although specific amino acid sequences described here areeffective, it is clear to those familiar with the art that amino acidscan be substituted or deleted without altering the effectiveness of thepeptides. Further, it is known that stabilization of the α-MSH sequencecan greatly increase the activity of the peptide and that substitutionof D-amino acid forms for L-forms can improve or decrease theeffectiveness of the peptides. For example, a stable analog of α-MSH,[Nle⁴, D-Phe⁷]-α-MSH, which is known to have marked biological activityon melanocytes and melanoma cells, is approximately ten times morepotent than the parent peptide in reducing fever. Further, adding aminoacids to the C-terminal of α-MSH(11-13) (SEQ. ID. NO. 1) sequence canreduce or enhance antipyretic potency. Addition of glycine to form the10-13 sequence (SEQ. ID. NO. 5) slightly decreased potency; the 9-13sequence (SEQ. ID. NO. 6) was almost devoid of activity, whereas thepotency of the 8-13 sequence (SEQ. ID. NO. 7) was greater than that ofthe 11-13 sequence (SEQ. ID. NO. 1). It is known that Ac-[D-K11]-α-MSH11-13-NH2 has the same general potency as the L-form of the tripeptideα-MSH (11-13) (SEQ. ID. NO. 1). However, substitution with D-proline inposition 12 of the tripeptide rendered it inactive. see e.g. Holdeman,M., et. al., Antipyretic Activity of a Potent α-MSH Analog, Peptides 6,273-5 (1985). Deeter, L. B., et. al., Antipyretic Properties ofCentrally Administered α-MSH Fragments in the Rabbit, Peptides 9, 1285-8(1989). Hiltz, M. E., Anti-inflammatory Activity of α-MSH (11-13)Analogs: Influences of Alterations in Stereochemistry, Peptides 12,767-71 (1991).

[0083] Biological functional equivalents can also be obtained bysubstitution of amino acids having similar hydropathic values. Thus, forexample, isoleucine and leucine, which have a hydropathic index +4.5and+3.8, respectively, can be substituted for valine, which has ahydropathic index of +4.2, and still obtain a protein having likebiological activity. Alternatively, at the other end of the scale,lysine (−3.9) can be substituted for arginine (−4.5), and so on. Ingeneral, it is believed that amino acids can be successfully substitutedwhere such amino acid has a hydropathic score of within about +/−1hydropathic index unit of the replaced amino acid.

[0084] Furthermore, these modified analogs of α-MSH and/or itsderivatives can also form dimers as exemplified by the KPV dimer in FIG.16.

EXAMPLE XII

[0085] A woman experiences discomfort in her vagina, vulva, and/orurinary tract. An examination by the physician can include a culture betaken from these area. After determining the cause of the discomfort,including any infection or inflammation, the physician may prescribe anantibiotic, anti-fungal, anti-viral, or anti-inflammation drug whereappropriate. In addition, the treatment can include a topicalapplication of pharmacologically effective amounts of α-MSH and/or itsderivatives carried in an ointment, cream, gel, dissolvable pill,aerosol spray, suppository, liquid solution for douche, or the absorbentmaterial of tampons. The topical treatment can be applied once ormultiple times according to the discretion of the physician until thecondition is resolved.

[0086] This example, where appropriate at the discretion of thephysician, can also be applied to a man who experiences discomfort inhis penis, testicles, and/or urinary tract. In addition, this topicalapplication of α-MSH and/or its derivatives may also be achieved withouta physician such as use as an over the counter drug.

[0087] The preceding Examples I-XII demonstrate the anti-infectionactivities and uses of α-MSH and/or its derivatives. These data areintended only as examples and are not intended to limit the invention tothese examples. It is understood that modifying the examples above doesnot depart from the spirit of the invention. It is further understoodthat the examples can be applied on its own or in combination with eachother.

0 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 8 <210> SEQ ID NO 1 <211>LENGTH: 3 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Designed polypeptide withanti-inflammatory, anti-microbial, anti-fungal, and anti-viralproperties. <400> SEQUENCE: 1 Lys Pro Val 1 <210> SEQ ID NO 2 <211>LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence <220>FEATURE: <223> OTHER INFORMATION: Designed polypeptide withanti-inflammatory, anti-microbial, anti-fungal, and anti-viralproperties. <400> SEQUENCE: 2 Met Glu His Phe Arg Trp Gly 1 5 <210> SEQID NO 3 <211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: Designed polypeptidewith anti-inflammatory, anti-microbial, anti-fungal, and anti-viralproperties. <400> SEQUENCE: 3 His Phe Arg Trp Gly Lys Pro Val 1 5 <210>SEQ ID NO 4 <211> LENGTH: 13 <212> TYPE: PRT <213> ORGANISM: ArtificialSequence <220> FEATURE: <223> OTHER INFORMATION: alpha-MSH withanti-inflammatory, anti-microbial, anti-fungal, and anti-viralproperties. <400> SEQUENCE: 4 Ser Tyr Ser Met Glu His Phe Arg Trp GlyLys Pro Val 1 5 10 <210> SEQ ID NO 5 <211> LENGTH: 4 <212> TYPE: PRT<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: Designed polypeptide with anti-inflammatory,anti-microbial, anti-fungal, and anti-viral properties. <400> SEQUENCE:5 Gly Lys Pro Val 1 <210> SEQ ID NO 6 <211> LENGTH: 5 <212> TYPE: PRT<213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHERINFORMATION: C-terminal sequences of alpha-MSH <400> SEQUENCE: 6 Trp GlyLys Pro Val 1 5 <210> SEQ ID NO 7 <211> LENGTH: 6 <212> TYPE: PRT <213>ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION:Designed polypeptide with anti-inflammatory, anti-microbial,anti-fungal, and anti-viral properties. <400> SEQUENCE: 7 Arg Trp GlyLys Pro Val 1 5 <210> SEQ ID NO 8 <211> LENGTH: 4 <212> TYPE: PRT <213>ORGANISM: Artificial Sequence <220> FEATURE: <221> NAME/KEY: DISULFID<222> LOCATION: From Cys to Cys forming a dimer with interchaindisulfide bond. <223> OTHER INFORMATION: Designed polypeptide withanti-inflammatory, anti-microbial, anti-fungal, and anti-viralproperties. <400> SEQUENCE: 8 Cys Lys Pro Val 1

1. A method of treating vaginitis comprising: administering atherapeutic amount of a polypeptide including an amino acid sequence KPVat C-terminus to an individual in need thereof.
 2. The method of claim 1wherein the polypeptide includes at least one amino acid in D-form. 3.The method of claim 1 wherein the amino acid sequence KPV is located ata C-terminal of the polypeptide.
 4. The method of claim 1 wherein thepolypeptide is N-acetylated or C-amidated or both.
 5. The method ofclaim 1 wherein the polypeptide is administered with a carrier.
 6. Themethod of claim 1 wherein the polypeptide is administered as a gel. 7.The method of claim 1 wherein the polypeptide is administered in a formselected from a group consisting of ointment, foam, balm, cream,dissolvable pill, aerosol spray, aerosol foam and liquid solution of adouche.
 8. The method of claim 1 wherein the polypeptide is administeredwith an applicator.
 9. The method of claim 8 wherein the applicator isselected from the group consisting of syringes, bandages, catheters, andspatulas.
 10. The method of claim 8 wherein at least a part of theapplicator is for insertion into a vagina.
 11. The method of claim 1wherein the vaginitis is caused by a bacterial infection.
 12. The methodof claim 11 wherein the bacterial infection is caused by Staphylococcusaureus.
 13. The method of claim 1 wherein the vaginitis is caused by afungal infection.
 14. The method of claim 13 wherein the fungus isCandida sp.
 15. The method of claim 14 wherein the fungus is Candidaalbicans.
 16. The method of claim 1 wherein the vaginitis is caused by aviral infection.
 17. A method of treating vaginitis comprising:administering a therapeutic amount of a dimer to an individual in needthereof wherein the dimer comprises KPV at C-terminus.
 18. The method ofclaim 17 wherein the dimer is a homodimer.
 19. The method of claim 17wherein the dimer is a heterodimer.
 20. The method of claim 17 whereinat least one amino acid is in D-form.
 21. The method of claim 17 whereinthe dimer is N-acetylated or C-amidated or both.
 22. The method of claim17 wherein the dimer comprises two monomers linked together by adisulfide bond and wherein each monomer comprises KPV.
 23. The method ofclaim 17 wherein the dimer comprises two monomers wherein each monomerhas a C-terminal Cysteine wherein the C-terminal Cysteine links themonomers together by a disulfide bond and wherein each monomer is KPV.24. The method of claim 17 wherein the dimer is administered with acarrier.
 25. The method of claim 17 wherein the dimer is administered inthe form of a gel.
 26. The method of claim 17 wherein the dimer isadministered in a form selected from a group consisting of ointment,foam, balm, cream, dissolvable pill, aerosol spray, aerosol foam andliquid solution of a douche.
 27. The method of claim 17 wherein thedimer is administered with an applicator.
 28. The method of claim 27wherein the applicator is selected from the group consisting ofsyringes, bandages, catheters, and spatulas.
 29. The method of claim 27wherein at least a part of the applicator is for insertion into avagina.
 30. The method of claim 17 wherein the vaginitis is caused by abacterial infection.
 31. The method of claim 30 wherein the bacterium isStaphylococcus aureus.
 32. The method of claim 17 wherein the vaginitisis caused by a fungal infection.
 33. The method of claim 32 wherein thefungus is from the genus Candida sp.
 34. The method of claim 33 whereinthe fungus is Candida albicans.
 35. The method of claim 17 wherein thevaginitis is caused by a viral infection.
 36. A method for treating ayeast infection comprising: administering a therapeutically effectiveamount of a composition to an area having a yeast infection wherein thecomposition comprises: a polypeptide having the amino acid sequence KPVat C-terminus; and a carrier.
 37. The method of claim 36 wherein theyeast infection is caused by Candida albicans.
 38. The method of claim36 wherein the carrier includes a gel.
 39. The method of claim 37wherein the composition further comprises an applicator.
 40. A methodfor treating a yeast infection comprising: administering to an areahaving the yeast infection a therapeutically effective amount of acomposition comprising: a dimer wherein the dimer comprises KPV atC-terminus; and a carrier.
 41. The method of claim 46 wherein thecarrier includes a gel.
 42. The method of claim 41 wherein thecomposition further comprises an applicator.
 43. A method for treating afungal, bacterial or viral infection comprising: administering to anarea having the infection, a therapeutically effective amount of acomposition comprising: a polypeptide having the amino acid sequence KPVat C-terminus; and a carrier.
 44. The method of claim 43 wherein thepolypeptide further comprises a dimer.
 45. The method of claim 43wherein the carrier includes a gel.
 46. The method of claim 40 whereinthe composition further comprises an applicator.